ABSTRACTω-3 PUFAs and polyphenols have multiple effects on inflammation in vivo and in vitro. The effects of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and resveratrol (RV) were investigated in LPS-stimulated peripheral blood leukocytes (PBLs) (i.e., acute inflammation) and IL-1β activated human chondrocytes (i.e., chronic inflammation). Inflammatory mediators including chemokines, cytokines, interleukins, and PGE2 were measured by multiplex analysis and gene expression was quantified by RT-PCR. In PBLs, RV decreased the secretion of PGE2, CCL5/RANTES, and CXCL8/IL-8 but increased IL-1β, IL-6, and IL-10. In contrast to RV, ω-3 PUFAs augmented the production of PGE2 and CXCL8/IL-8. EPA and DHA similarly affected the pattern of inflammatory mediators. Combination of RV and ω-3 PUFAs exerted synergistic effects on CCL5/RANTES and had additive effects on IL-6 or CXCL8/IL-8. Both ω-3 PUFAs and RV reduced catabolic gene expression (e.g., MMPs, ADAMTS-4, IL-1β, and IL-6) in activated chondrocytes. The data suggest that ω-3 PUFAs and RV differ in the regulation of acute inflammation of peripheral blood leukocytes but have common properties in modulating features related to chronic inflammation of chondrocytes.

fig3: Effect of combinations of substances on inflammatory mediators produced by activated PBLs. PBLs were cultured for 24 h with or without the indicated substances and their combinations. In the case of inhibition, the combination index (CI) was calculated and is indicated in the figure. Dashed lines in the bar graphs indicate the computed sum of the respective single substance treatments.

Mentions:
Since we observed similar and opposite effects of substances, we investigated the pattern on inflammatory parameters produced when PBLs were treated with a combination of substances. To this aim, cells were activated in the presence of different concentrations and ratios of individual substances and the secreted mediators determined (Figure 3). PGE2 production was dominated by the inhibitory effect of RV, which partially counterbalanced the enhancing effect of DHA (Figure 3). CXCL8/IL-8 production was controlled by DHA, since the combined treatment with RV did not result in an intermediate production. Combinations of RV and DHA synergistically inhibited CCL5/RANTES secretion, as computed by the Chou-Talalay algorithm (Figure 3). Both RV and DHA concentration-dependently enhanced IL-6 secretion and combinations thereof had additive effects. IL-1β, however, appeared to be synergistically enhanced by RV and DHA, since the effect of combined substances largely exceeded the sum of RV and DHA applied individually.

fig3: Effect of combinations of substances on inflammatory mediators produced by activated PBLs. PBLs were cultured for 24 h with or without the indicated substances and their combinations. In the case of inhibition, the combination index (CI) was calculated and is indicated in the figure. Dashed lines in the bar graphs indicate the computed sum of the respective single substance treatments.

Mentions:
Since we observed similar and opposite effects of substances, we investigated the pattern on inflammatory parameters produced when PBLs were treated with a combination of substances. To this aim, cells were activated in the presence of different concentrations and ratios of individual substances and the secreted mediators determined (Figure 3). PGE2 production was dominated by the inhibitory effect of RV, which partially counterbalanced the enhancing effect of DHA (Figure 3). CXCL8/IL-8 production was controlled by DHA, since the combined treatment with RV did not result in an intermediate production. Combinations of RV and DHA synergistically inhibited CCL5/RANTES secretion, as computed by the Chou-Talalay algorithm (Figure 3). Both RV and DHA concentration-dependently enhanced IL-6 secretion and combinations thereof had additive effects. IL-1β, however, appeared to be synergistically enhanced by RV and DHA, since the effect of combined substances largely exceeded the sum of RV and DHA applied individually.

Bottom Line:
ω-3 PUFAs and polyphenols have multiple effects on inflammation in vivo and in vitro.Combination of RV and ω-3 PUFAs exerted synergistic effects on CCL5/RANTES and had additive effects on IL-6 or CXCL8/IL-8.Both ω-3 PUFAs and RV reduced catabolic gene expression (e.g., MMPs, ADAMTS-4, IL-1β, and IL-6) in activated chondrocytes.

ABSTRACTω-3 PUFAs and polyphenols have multiple effects on inflammation in vivo and in vitro. The effects of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and resveratrol (RV) were investigated in LPS-stimulated peripheral blood leukocytes (PBLs) (i.e., acute inflammation) and IL-1β activated human chondrocytes (i.e., chronic inflammation). Inflammatory mediators including chemokines, cytokines, interleukins, and PGE2 were measured by multiplex analysis and gene expression was quantified by RT-PCR. In PBLs, RV decreased the secretion of PGE2, CCL5/RANTES, and CXCL8/IL-8 but increased IL-1β, IL-6, and IL-10. In contrast to RV, ω-3 PUFAs augmented the production of PGE2 and CXCL8/IL-8. EPA and DHA similarly affected the pattern of inflammatory mediators. Combination of RV and ω-3 PUFAs exerted synergistic effects on CCL5/RANTES and had additive effects on IL-6 or CXCL8/IL-8. Both ω-3 PUFAs and RV reduced catabolic gene expression (e.g., MMPs, ADAMTS-4, IL-1β, and IL-6) in activated chondrocytes. The data suggest that ω-3 PUFAs and RV differ in the regulation of acute inflammation of peripheral blood leukocytes but have common properties in modulating features related to chronic inflammation of chondrocytes.